BIO-INORGANIC CHEMISTRY (CHEM.

411) (3 Credit Hours)

COURSE OUTLINE

Essential and non-essential elements in biological systems. Surveys of functions, uses and
toxicity. The porphyrin ring system and matalloporphyrins in nature. Heavy metals in biological
systems; health effects. Chemical toxicology; a brief survey. Chemical carcinogens, harmful
drugs.
LECTURE NOTE 1.

BIO-INORGANIC CHEMISTRY (CHEM. 411) (3 Credit Hours)

TERMS RELATED TO BIO-INORGANIC CHEMISTRY

1. Active center: Location in an enzyme where the specific reaction takes place

2. Allosteric enzyme: This can bind a small regulatory molecule that influences catalytic activity

3. Apo-enzyme: This is an enzyme that lacks its metal center or prosthetic groups

4. Bio-membrane: Sheet like assemblies of proteins and lipids (bilayer)

5. Calmodulin: Ca binding protein involved in metabolic regulation

6. Carboanhydrase: Zn-containing enzyme that catalyzes the reversible decomposition of

carbonic acid to carbon dioxide and water

7. Charge-transfer complex: An aggregate of two or more molecules in which charge is

transferred from a donor to an acceptor.

8. Chlorin: 2, 3-Dihydroporphyrin, reduced porphyrin with two non-fused saturated carbon

atoms (C-2, C-3) in one of the pyrrole rings.

9. Chlorophyll: Magnesium complex of a porphyrin in which a double bond in one of the pyrrole
rings has been reduced. A fused cyclopentanone ring is also present

10. Cluster: Metal centers grouped close together which can have direct metal bonding or
through a bridging ligand, e.g. ferredoxin

11. Cobalamin: Vitamin B12, substituted corrin-Co (III) complex

12. Coenzyme: A low-molecular-weight, non-protein organic compound (often a nucleotide)

participating in enzymatic reactions

13. Cofactor: An organic molecule or ion (usually a metal ion) that is required by an enzyme for
its activity. It may be attached either loosely (coenzyme) or tightly (prosthetic group).

14. Cooperativity: The phenomenon that binding of an effector molecule to a biological system
either enhances or diminishes the binding of a successive molecules, e.g. hemoglobin

15. Corrin: Ring-contracted porphyrinderivative that is missing a carbon

16. Cytochrome: Hemeprotein that transfers electrons, and exhibits intense absorption bands.
The iron undergoes oxidation-reduction between oxidation states Fe(II) and Fe(III).
17. Cytochrome-c oxidase: The major respiratory protein of animal and plant mitochondria. It
catalyzes the oxidation of Fe(II)-cytochrome c, and the reduction of dioxygen to water. Contains
two hemes and three copper atoms, arranged in three centers.

18. Cytochrome: General term for a group of heme-containing monooxygenases. The reaction
with dioxygen appears to involve higher oxidation states of iron.

19. Cytoplasm: The part of protoplasm in a cell outside of and surrounding the nucleus

20. Dehydrogenase: An oxidoreductase which catalyzes the removal of hydrogen

21. Desferrioxamine(dfo): Chelating agent used world-wide in the treatment of iron overload
conditions, such as hemochromatosis and thalassemia.

22. Dismutase: Enzyme that catalyzes a disproportionation reaction

23. Entatic state: A state of an atom or group which has its geometric or electronic condition
adapted for function. Derived from entasis(Greek) meaning tension

24. Enzyme: A macromolecule that functions as a biocatalyst by increasing the reaction rate

25. Ferredoxin: A protein containing more than one iron and acid-labile sulfur, that displays
electron-transferactivity but not classical enzyme function

26. Ferritin: An iron storage protein consisting of a shell of 24 protein subunits, encapsulating
up to 4500 iron atoms in the form of a hydrated iron(III) oxide.

27. Heme: A near-planar coordination complex obtained from iron and dianionic porphyrin

28. Hemerythrin: A dioxygen-carrying protein from marine invertebrates, containing an oxo-

bridged dinuclear iron center

29. Hemocyanin: A dioxygen-carrying protein (from invertebrates, e.g arthropods and molluscs),
containing dinuclear type 3 copper sites

30. Hemoglobin: A dioxygen-carrying hemeprotein of red blood cells

31. Ion channel: Enable ions to flow rapidly through membranes in a thermodynamically
downhill direction after an electrical or chemical impulse. Their structures usually consist of 4-6
membrane-spanning domains. This number determines the size of the pore and thus the size of
the ion to be transported

32. Ionophore: A compound which can carry specific ions through membranes

33. Ion pumps: Enable ions to flow through membranes in a thermodynamically uphill direction
by the use of an energy source. They open and close upon the binding and subsequent hydrolysis
of ATP, usually transporting more than one ion towards the outside or the inside of the
membrane

34. Metalloenzyme: An enzyme that, in the active state, contains one or more metal ions

35. Mitochondria: This is Cytoplasmic organelles, produce ATP by oxidative phosphorylation

36. Model: A synthetic coordination entity that closely approaches the properties of a metal ion
in a protein and yields useful information concerning biological structure and function

37. Myoglobin: A monomeric dioxygen-binding hemeprotein of muscle tissue, structurally

similar to a subunit of hemoglobin

38. Nucleic acids: Macromolecules composed of sequences of nucleotides that perform several
functions in living cells, e.g. the storage of genetic information.

39. Nucleosides: Compounds in which a purine or pyrimidine base is beta-N-glycosidically

bound to C-1 of either 2-deoxy-D-ribose or of D-ribose, but without any phosphate groups

40. Nucleotides: Nucleosides with one or more phosphate groups esterified mainly to the 3'-or
the 5'-position of the sugar moiety.

41. Photosynthesis: A metabolic process in plants and certain bacteria, using light energy
absorbed by chlorophyll and other photosynthetic pigments for the reduction of CO2, followed
by the formation of organic compounds.

42. Plastocyanin: An electron transfer protein, containing a type 1 copper site, involved in plant
and cyanobacterial photosynthesis, which transfers electrons to Photosystem1

43. Rieske protein: An iron-sulfur protein of the mitochondrial respiratory chain, containing a
[2Fe-2S] cluster.

44. Rubredoxin: An single iron-sulfur protein, function as an electron carrier.

45. Soret band: Strong absorption band in the blue region of the optical absorption spectrum of a
hemeprotein

46. Substrates: A compound that is transformed under the influence of a catalyst.

47. Trace elements: Elements required for physiological functions in very small amounts, e.g.
Co, Cu, F, Fe, I, Mn, Mo, Ni, Se, V, W, and Zn
THE ROLE OF ESSENTIAL AND NON-ESSENTIAL ELEMENTS IN BIOLOGICAL
SYSTEMS

If we look at the periodic table we can find around 25 and others elements that are required by
most, if not all, biological systems and these are given below:

Hydrogen

Hydrogen is extremely important in biology. It can be incorporated into covalent bonds with
many non-metals, such as carbon and nitrogen, notably by the action of light. It can be
transferred in an important number of biological redox reactions involving one or two electron
transfers, and it can participate in the generation of the proton gradients across biological
membranes, which are universally used for ATP synthesis.

Helium
Helium, like the other members of its family, is an inert gas often used in balloons on account of
its low density, and when inhaled results in a comic transposition of the human voice to a
significantly higher register.

Lithium
Lithium, while not required for life, is used therapeutically in the form of lithium carbonate for
the treatment of manic depression; although its mechanism of action remains a mystery.
Effective treatment requires attaining serum lithium concentrations of between 0.8 and 1.2
mol/L.

Boron
Boron is an essential trace element for plants, and may well turn out to be essential for mammals
as well. The boron-containing polyether–macrolide antibiotic, boromycin, was isolated as a
potential anti-HIV agent.

Fluorine
The addition of fluoride in drinking water to retard dental caries, particularly in children, has
been criticized on the grounds of potential toxicity, but the concentrations used are many orders
of magnitude below that which would be required to inhibit enzymes such as enolase in the
glycolytic pathway. The key enzyme of DNA synthesis, thymidylate synthase, is inhibited by the
anti-tumour drug 5-fluorodeoxythymidylate a so-called ‘suicide substrate’, because it inhibits the
enzyme only after undergoing part of its normal catalytic reaction.

Sodium
Sodium is involved in ionic gradients and in osmotic regulation, and, despite its much higher
extracellular concentration, has to be kept out of many cells by the action of an energy
consuming Na/KATPase.

Magnesium
Magnesium has its role intimately intertwined with phosphate: in many phosphoryl transfer
reactions, as Mg-ATP in muscle contraction, in the stabilization of nucleic acid structures as well
as in the catalytic activity of ribozymes (catalytic RNA molecules). It also serves as a structural
component of enzymes, and is found as the metal center in chlorophylls, which absorbs light
energy in photosynthesis

Carbon

Living thing used carbon in a vast number of molecules in their bodies, carbohydrates, lipids,
proteins, and nucleic acid all contain carbon thus this means that we found carbon in everything
from cell membrane to hormones to DNA. Carbon’s versatility means that it is the basis of many
of the complex organic compounds vital to life and the backbone of polypeptide chain that form
different protein in the body of living thing.

Oxygen

Oxygen is very essential in mammals cellular respiration is the term used to describe the phase of
the digestive process when food breaks down to supply cells. During cellular respiration, cell
used oxygen to break down sugar to produce ATP (Adenosine Triphosphate). ATP is a molecule
that supply cells with energy. The by-product of the process are CO2 and H2O

Nitrogen

Nitrogen is a component of amino acid and urea. AAs are the building block of all protein.
Proteins comprises not only structural component such as muscles, tissue and organ but also
enzymes and hormones essential for the functioning of all living thing. Urea is a by-product of
protein digestion. The nitrogen in protein and urea is organic nitrogen. We use the term organic
nitrogen to describe a nitrogen compound that had its origin in living materials.

Aluminium

Aluminium while extremely abundant in the earth’s crust, is not used by living organisms: it is a
notorious neurotoxin, but its involvement as a cause of Alzheimer’s disease seems less likely
than was thought a few years ago. It is clear that acid rain, due to sulfur dioxide and nitrogen
oxide emissions, increases the solubility and hence the bioavailability of aluminium. In the
forests on the mountain slopes, the pH values reached below 3, with disastrous effects on the tree
population. Another effect of acid rain could have been to change the usual association of
aluminium in the soil with silicate (predominant above pH 6.5) for phosphate, rendering
aluminium more toxic. This may be the reason why silicon is essential, namely that it keeps
aluminium in a non-toxic form as aluminium silicate. While silicon is required as a trace element
in most animals, in plants, particularly grasses, and in many unicellular organisms, such as
diatoms, it is a major structural element. The importance of phosphorus and sulfur is obvious, the
latter often associated with iron in an important family of proteins that contains iron–sulfur
clusters.

Chlorine

This is another essential element in large part because, with all of the positively charged metal
ions around, anions are obviously required for charge neutralization. One of the most common
genetic disorders in man, cystic fibrosis (often referred to as mucoviscidosis, because of the
viscous nature of bronchial secretions, resulting in frequent respiratory infections), is due to the
production by epithelial cells that line the lungs, digestive tract, sweat glands and genitourinary
system, of a defective form of a protein called cystic fibrosis transmembrane conductance
regulator (CFTR), which is a chloride channel. Detailed structural analysis of closed and open
conformations of bacterial chloride channels has shown that they can be closed by a glutamate
residue, which replaces a third Cl- ion on the extracellular site of the channel. In the closed
conformation, the ion-binding sites Sint and Scen are occupied by Cl- ions and the ion-binding site
Sext is occupied by the side chain of Glu 148, whereas in the opened conformation Glu 148 has
moved out and the binding site is occupied by a third Cl - anion. Argon, an inert gas, has the
useful property of being heavier than air, thus making it the ideal medium in which to work
anaerobically (keep everything at the bottom of your argon-flushed glove box!).

Potassium

Potassium, like sodium, is involved in ionic equilibria, and the opening and closing of sodium
and potassium ion channels create the electrochemical gradients across cell membranes that
transmit nerve impulses and other information and regulate cellular function.

Calcium

Calcium, a crucial second messenger signaling key changes in cellular metabolism, is also
important in muscle activation, in the activation of many proteases, both intra- and extracellular,
and as a major component of a range of biominerals, including bone.

Cobalt

This is an important element in the formation of Cobalamin or Vitamin B 12, it is not easily
assimilated in the body and is stored in Red Blood Cell (RBC), liver, plasma, kidney and
pancreas. It promote the formation of RBC. Activate enzymes, replaces zinc in some enzymes,
Cobalt salt in in small amount are essential to many life forms including human. It is at the core
of a vitamin called B12. Moreover, it is essential for the formation of hemoglobin and aid in
normal growth and appetite.

Vanadium

Vanadium is known to be essential, and is a constituent of some haloperoxidases as well as

nitrogenases in some nitrogen-fixing organisms. It is particularly abundant in tunicates (a species
of marine organisms) and in Amanita toadstools.

Chromium

Chromium presents a big enigma: it appears to be essential for man, yet we do not have a clue as
to what it might do—indeed most chromium compounds are toxic. The biologically relevant
form, trivalent Cr3+, is required for carbohydrate and lipid metabolism in mammals. Chromium
has become extremely popular as a nutritional supplement, weight loss and muscle-development
agent, second only to Ca-containing products among mineral supplements.
Manganese

Manganese is essential for man, although possibly its most significant contribution to biology is
its incredible chemistry as a tetra-manganese centre in the splitting of water by photosystem II in
plants, and originally at a much earlier point in geological time, in cyanobacteria. This reaction
generates oxygen, which of course changed the whole pattern of the evolution of planet Earth.
Perhaps this was the greatest pollution event in the history of our planet, which progressively
moved us from an essentially reducing atmosphere to the oxidative world that we now know.
There were, of course, advantages—respiration is almost 20 times more effective at producing
ATP than fermentation. However, the oxygen paradox, as it has been termed, also resulted in the
production of toxic reactive oxygen species, notably the hydroxyl radical through the well-
known Fenton reaction. Another consequence of the appearance of dioxygen was that divalent
copper became much more bioavailable, whereas trivalent iron underwent hydrolysis,
polymerization and precipitation, making it much more difficult to extract.

Iron

Iron is essential for almost all living organisms, most probably because of its role in forming the
amino acid radicals required for the conversion of ribonucleotides to deoxy ribonucleotides in
the Fe-dependent ribonucleotide reductases. In those organisms, such as Lactobacilli, which do
not have access to iron, their ribonucleotide reductases use a cobalt-based cofactor, related to
vitamin B12.

Copper

Copper, like iron, is frequently encountered in reactions involving dioxygen. The copper enzyme
laccase catalyses the oxidation of uroshiol (the same poisonous substance found in poison oak
and ivy) in the production of Japanese lacquer. It is the products of uroshiol oxidation, which are
responsible for the lacquer’s remarkable material properties.

Gallium

Gallium is non-essential, but on account of the similarity between Ga 3+ and Fe3+ it binds to iron
transport and storage proteins such as transferrin and ferritin. The radioactive isotope of
gallium,Ga, concentrates to a large extent in many tumours and at sites of inflammation and
infection, and since many tumours overexpress the transferring receptor it can be used for
tumour imaging.
Arsenic

Arsenic is highly toxic, and indeed much speculation has surrounded arsenic poisoning as the
cause of death. Arsenic trioxide has been approved by the Food and Drug Administration (FDA)
of the USA for the treatment of acute promyelocytic anaemia in adult patients who fail to
respond to other chemotherapy, or have relapsed disease.

Selenium

Selenium is essential for many species including man, on account of its presence in a number of
enzymes, notably glutathione peroxidase, an important antioxidant enzyme. It is incorporated
into selenoenzymes in the form of selenocysteine.

Bromine

Bromine is thought to be essential for plants and animals, although no known biological role has
been established. It has flame-extinguishing characteristics and is used in fireproofing agents and
to make flame-resistant plastics.

Molybdenum

Molybdenum is essential for a number of enzymes, for example xanthine oxidases in mammals
and nitrogenases in nitrogen-fixing bacteria.

Tin

Tin is thought to be an essential trace element for some species, although its precise role remains
unknown. Some therapeutic uses of tin compounds have been proposed, and triorganotin
carboxylates are effective bacteriocides and pesticides. Tin is, of course, an important component
of a number of alloys, with copper in bronze (the Bronze Age began about 3500 BC), and with
lead in pewter.

Iodine

Iodine is an essential element with an important role in mammals in the regulation of

metabolism, through the action of the two related hormones triiodothyronine (T 3) and thyroxine
(T4) produced by the thyroid gland. The biosynthesis of these two hormones occurs through the
iodination, rearrangement and subsequent hydrolysis (proteolysis) of tyrosine residues in the
thyroglobulin protein. Iodine, which is relatively scarce, is actively concentrated in the thyroid
gland where both T3 and T4 are produced.
Tungsten

Tungsten, still considered to be essential for some organisms, is required as a cofactor in a

number of prokaryotic enzymes. It was also the element that would replace osmium and tantalum
(and before them, carbon) in the electric lamps of the early twentieth century, which gradually
replaced gas lamps

Zinc

Zinc is found in more than 300 enzymes, where it plays both a catalytic and a structural role. It is
the only metal to have representatives in each of the six fundamental classes of enzymes
recognized by the International Union of Biochemistry: oxidoreductases, such as alcohol
dehydrogenase and superoxide dismutase; transferases, such as RNA polymerase and aspartate
transcarbamoylase; hydrolases, such as carboxypeptidase A and thermolysin; lyases, such as
carbonic anhydrase and fructose-1,6-bisphosphate aldolase; isomerases, such as
phosphomannose isomerase; and ligases, such as pyruvate carboxylase and amino acyl-tRNA
synthetases.

Not only is zinc involved in enzymes, where it plays both a catalytic and a structural role, but
there are also growing numbers of nucleic acid-binding proteins with essential Zn atoms. This
demonstrates that Zn is also widely involved in the regulation of the transcription and translation
of the genetic message. The bioinorganic chemistry of zinc is dominated by a number of factors,
the most pertinent of which are: The divalent zinc ion is redox inactive, in contrast, for example,
to manganese, iron and copper. Its d10 configuration means that not only does it have no d-d
transitions, and therefore no absorption spectroscopy, but also its complexes are not subject to
ligand field stabilization effects such that Zn has no ligand field constraints on its coordination
geometry. Coordination number and geometry are therefore dictated only by ligand size and
charge. This means that zinc can, in principle, adopt highly flexible coordination geometry.
However, in most zinc proteins there is a strong preference for tetrahedral coordination,
frequently slightly distorted, which enhances both the Lewis acidity of the zinc center and the
acidity of a coordinated water molecule. Only Cu (II) is a better Lewis acid.

A few cases of zinc in five-coordinate distorted trigonal bipyramidal geometry have been
reported. Since zinc is of borderline hardness, it can bind oxygen (Asp, Gu, H 2O), nitrogen (His)
and sulfur (Cys) ligands. Three types of zinc-binding sites have been recognized in zinc enzymes
(catalytic sites, structural sites and cocatalytic sites). Many of these zinc enzymes are peptidases
and amidases, involved in the cleavage of amide bonds—they include peptidases, such as
thermolysin and carboxy-peptidases; -lactamases, which destroy the four-member -lactam rings
in penicillins; and matrix metalloproteinases, which degrade extracellular matrix components
such as collagen.
Zinc enzymes also participate in the cleavage of the phosphodiester bonds in both DNA and
RNA, and their role extends beyond catalysis of hydrolytic reactions to include the important
lyase, carbonic anhydrase and the oxidoreductase, alcohol dehydrogenase. Some of the zinc-
based motifs found in proteins involved in the regulation of nucleic acid and protein synthesis.
LECTURE NOTE 2

BIO-INORGANIC CHEMISTRY (CHEM. 411) (3 Credit Hours)

THE PORPHYRIN RING SYSTEM AND METALLOPORPHYRINS IN NATURE

Porphyrins are one of the vital chemical units essential for several life processes on the earth.
Many biological molecules function with prosthetic groups essentially made of these units.
Chlorophylls of chloroplasts which drive photosynthesis, heme as a component of hemoglobin
that transports oxygen to animal tissues and as the central unit of myoglobin ensures the storage
of oxygen - all these have active sites essentially made of porphyrin core'". Over the years, a
great deal of concerted efforts have brought to light substantial understanding of the structure-
function relationship in these natural porphyrins

A large variety of synthetic porphyrins and their metalloderivatives were made over the years to
study the porphyrin based natural systems. The search for anti-cancer drugs, useful catalysts,
semiconductors and superconductors, electronic materials with novel properties has also made
this synthetic porphyrin chemistry a very actively probed one by chemists, biologists and
physicists alike. The synthetic meso-substituted porphyrins offer a great advantage to study the
physical and chemical properties of the porphyrin nucleus quantitatively by a judicious choice of
the substituents that may be attached on the periphery. Metalloporphyrins are widely and
intensely investigated in the area of catalysis and also as models and mimics of enzymes lie
catalase, peroxidases, P450 cytochromes or as transmembrane electron transport agents. They
have also been used as NMR image enhancement agents, Nonlinear optical materials" and DNA-
binding or cleavage agent". Currently there is interest in using chelated radioactive metal
isotopes as diagnostic imaging and therapeutic agents. In that context porphyrins are excellent
compounds because of their extremely high stable constant with many metal ions.

The Porphyrin System

Porphyrins are basically cyclic tetrapyrrole derivatives with a highly delocalised planar
framework having a core structure. They can exist in varied forms by having different peripheral
substituents at all the eight pyrrole P-carbon atom and the four meso-carbon centres and also by
undergoing certain structural variations.
Porphyrin is an 18- π: electron system and hence exhibits aromaticity. The simplest porphyrin is
known as porphine which is the H-analogue (R1 - R12 = H) of 1. Besides many synthetic and
naturally occurring ones with the core structure mentioned above, there are many biologically
active systems which also have porphyrin-like structures which are given in

Porphin Chlorin Phorbin

 Bacteriochlorin Porphyrazine Phthalocyanine

The porphyrin ring provides a vacant site at its center, ideally suited for metal incorporation. The
NH protons inside the ring of porphyrins possess acidic character and hence can get deprotonated
to give porphyrinato ions. These dianion species with their electronically sensitive planar π
framework and central cavity with more or less rigid size exhibit remarkable ligation
characteristic towards metal ions. Thus derivatives of porphyrins with almost all metals and
semi-metals have been synthesized. A crucial factor to form stable metalloporphyrins seems to
be the compatibility of porphyrin ring size with the ionic radii of the metal cations". Hence stable
complexes generally result when these two sizes match while their instability tends to increase
when the size of the cation is too big or too small with very few exceptions. The porphyrinato
dianion is ideally suited to act as a tetradenate ligand with metal ions. Thus the minimum
coordination number of the metal ion possible in a metalloporphyrin is four. A size matching
divalent metal ion would give neutral complex, while a higher valent cation would carry with it
balancing anion (s) mostly covalently bound with the metal, in addition to the porphyrinato ion.
The normal coordination geometry around the metal ion in the former species would be square
planar, while in the latter case the coordination geometry would be square pyramidal.
Coordination number greater than four is also possible. The two ligands of the six coordinate
metalloporphyrins are found on the opposite sides of the porphyrinato plane yielding complexes
with tetragonal or octahedral geometries. Ability to exhibit variable oxidation states of metals in
their metalloporphyrins is another important feature in this class of compounds. They are also
capable of stabilizing metal ions in their unusual oxidation states, which have resulted in
extensive studies revealing interesting chemistry.
Electronic Properties of Porphyrins and Metalloporphyrins

The most useful spectroscopic technique for the study of porphyrin and their metalloderivatives
is the electronic absorption spectroscopy. As the spectral absorptions are found to be sensitive to
the nature of porphyrins and its surroundings, vital information could be obtained on the nature
of chemical environment in which they exist and on the role these molecules play in key
biological functions they take part, all by just monitoring the electronic spectra in respective
conditions. Since the present study deals with some aspects of aggregation characteristics of
porphyrins and environment effects on their electronic spectra provide a vital tool to study them.
A brief description on the origin of the spectra of the porphyrins and their metalloderivatives are
given below.

The electronic heart of a porphyrin is the inner 16-membered ring with 18-π electrons. The ring
is structured with basic fourfold symmetry, including four nitrogen atoms directed towards the
center. This electronic heart is responsible for the unique porphyrin-type optical spectra, which
are then perturbed to a greater or lesser extent by various chemical modifications to the basic
structure.

Porphyrin and its metal derivatives are of considerable spectroscopic interest because of their
simplicity and uniqueness. The optical absorptions of porphyrin are determined essentially by the
π - electrons on the porphyrin ring, with only minor perturbation from the electrons of the central
substituents. The optical absorption spectra is an important spectral phenomenon to distinguish
between the free-base porphyrins and their metalloderivatives. The spectrum changes from four-
banded to a two-banded spectrum on metalation.

Types of Porphyrins

There are a variety of porphyrins that are of special significance. Described below in brief are
some of them.

1. Protoporphyrin

Protoporphyrin (9) contains four methyl groups, two vinyl groups and two propionic acid groups
on its periphery. Fifteen different isomeric protoporphyrins differing in the sequence of
substitution of the above groups in eight available positions are possible. Hemin is the prosthetic
group of hemoglobin, myoglobin, peroxidase, P-type cytochromes, catalase, tryptophan
pyrrolase, cytochrome P450 and many other proteins. Synthesis of porphyrins derived from those
found in animal materials invariably involves hemin or protoporphyrin as starting material.

Protoporphyrin dimethyl ester is most easily obtained from hemin by iron removal and
esterification. The original Grinstein procedure that accomplishes both operations simultaneously
involves passage of gaseous HCI through a methanolic solution of hemin
2. Deuteroporphyrin

Deuterohemin (10) was a key intermediate in Fischer's total synthesis of hemin. A Friedel-Crafts
acylation of deuterohemin followed by reduction and 26.27 dehydration gave protohemin.
Because they have two free P-positions, deuteroporphyrin and deuterohemin have been used as
starting materials in the synthesis of several porphyrins in which the vinyl groups of
protoporphyrin are replaced by other functionalities. The 2,4dibromo, diacetyl, dipropionyl and
dihydroxymethyl derivatives are all synthesised directly from deuteroporphyrin. Purification of
deuterohemin was done using deuteroporphyrin in Caughey's methodz8. The iron chelate is
obtained by reinsertion of iron into deuteroporphyrin dimethylester. Deuteroporphyrin-free acid
may be obtained from the dimethyl ester by hydrolysis, or from purified deuterohemin free acid
by iron removal.
3. Hematoporphyrin

Hematoporphyrin (11) was the fmt porphyrin isolated from natural materials. It was prepared by
Thudichum in 1867 by the sulfuric acid treatment of blood. Hematoporphyrin can also be
prepared by the treatment of hemin with HBr in acetic acid. The HBr adduct is then decomposed
with water to give hematoporphyrin. Hematoporphyrin is by far the most labile natural
porphyrins commonly used in the laboratory

4. Mesoporphyrin

Mesoporphyrin (12) is used in biological and chemical studies especially where the liability of
the vinyl groups of protoporphyrin is a factor. The classical method of mesoporphyrin synthesis
was HI reduction of protohemin Better result was obtained by the catalytic reduction over PdO
of protohemin, protoporphyrin or protoporphyrin dimethyl ester in formic acid at elevated
temperature. The most convenient method for obtaining large amount of mesoporphyrin was
developed by caughey. In this method hydrogen was bubbled through a formic acid solution of
hemin over PdO
5. Diacetyldeuteroporphyrin

This prophyrin (13) possesses electron withdrawing acetyl groups in the 2- and 4-positions.
Fischer's classical method is now used for the synthesis of diacetyldeuteroporphyrin. Crude
deuterohemin was dissolved in acetic anhydride at 0°C followed by the addition of anhydrous
SnCl4 under constant stirring. Longer reaction times result in substantial decomposition, and
shorter times give substantial amount of the monoacetylated product. The solution is further
acidified using 0.1N HCI and thoroughly stirred to avoid acetic anhydride formation. The crude
diacetyldeuterohemin is isolated by suction filtration.
6. Diformyldeuteroporphyrin

Diformyldeuteroporphyrin (14) with two strongly electron-withdrawing formyl groups has been
used in the studies of the relationship of heme structure to heme protein function3'. Fischer's
method, alkylation of deuterohemin with dichloromethyl methyl ether in the presence of SnBr4
followed by hydrolysis, gives a mixture of products including the monoformyl compounds.

7. Porphyrin C

C-type cytochromes (15) contains a prosthetic group, a covalent bound hemin, that can be
regarded as been formed by the addition of two cystane sulfhydryls across the double bonds of
protoheme. Treatment of cytochrome C with silver or mercury salts result in the formation of
optically active hematoporphyrin. Acid hydrolysis of horse heart cytochrome C results in the
formation of a homogenous amphoteric porphyrin fraction, which has been shown to be
porphyrin.

Porphyrins in Life Chemistry

Metal complexes of porphyrins and related compounds are important prosthetic groups that
assemble to form a wide variety of proteins and enzymes working as redox and rearrangement
catalysts. The nature being a master designer, has chosen and retained the best choice of
molecular assemblies in living systems to carry out very specifically the functions they are
intended for and also to achieve them with the maximum efficiency.

A large number of naturally occurring porphyrin have been isolated and characterized. Of these
protoporphyrins are the most abundant and widely characterized ones. It is found in hemoglobin,
myoglobin, heme enzymes and most of the cytochromes.
The biological and chemical importance of metalloporphyrins has brought to focus intense
interest in the nature of the metal ligand linkages in such complexes as well as all the
physicochemical properties of the macrocycles. The macrocycle has the ability to function as a
reservoir of electrons and control the reactivity at the axial position of metal, which usually
serves as a catalytic site in heme enzyme. Because of their ubiquitousness and the variety of their
natural functions, heme proteins have been investigated on multi- and interdisciplinary levels.
These proteins all containing an iron porphyrin as the prosthetic group, are responsible for
oxygen transport and storage (hemoglobin and myoglobin), electron transport (cytochromes),
oxygen reduction (cytochrome oxidase), hydrogen peroxide utilization and destruction
(peroxidases and catalases), and hydrocarbon oxidation (Cytochrome).

Also the various functions of heme proteins in the transport, storage and reactions with dioxygen
are made possible by different and selective interactions of diverse proteins with the heme
groups'0. These differences are brought about largely by the axial ligands provided by ancillary
groups of the protein and from the nature of the pockets on either side of the porphyrin.
Important porphyrin based natural systems are the hemoglobin, myoglobin, chlorophyll, heme
enzymes and the cytochromes.

1. Hemoglobin and myoglobin

Hemoglobin and myoglobin are high molecular weight protein systems containing iron
(I1) protoporphyrin IX units. They are responsible for oxygen transport and storage in
higher animals9. Hemoglobin transport dioxygen from its source to the site of use inside
the muscle cells. There the oxygen is transferred to myoglobin for use in respiration.
Myoglobin which is responsible for storing oxygen in cells, has high affmity for the
dioxygen even at low partial pressure but in lungs, hemoglobin takes up high amount of
oxygen at high partial pressure. This special property of hemoglobin is attributed to the
'cooperative effect" caused by the decrement in size of Fe 2+ in central hole due to spin
change (from high spin to low spin) on dioxygen complexation.
The iron in hemoglobin and myoglobin is in the ferrous state and has to have an N-base
(histidine) coordinated to the metal from one of the sides of the plane to have dioxygen
bound at the vacant sixth coordination site. The oxidized form of iron, ie. Ferric state,
called metmyoglobin and methemoglobin will not bind oxygen. The free heme is
immediately oxidized in the presence of oxygen and water and thus renders useless for
Oz transport.

2. Chlorophyll
Chlorophyll is the green colouring matter of leaves and green stems, and its presence is
essential for photosynthesis. In green plants it is the chlorophyll which absorbs the light
energy. Chlorophyll is basically magnesium derivative of porphyrins and some slight
structural changes in porphyrin moiety results in different classes of chlorophyll with
slight difference in photocatalytic properties. All of the chlorophylls absorb light very
intensely, particularly at relatively long wavelength regions43. The light energy absorbed
by a chlorophyll molecule become delocalized and spread throughout the entire
electronic structure of the excited molecule.
The photosynthetic pigments in the chloroplasts of plants consists of two functional units
namely photosystem I and photosystem 11. Photosystem I contains chlorophyll, β-
carotene and a single molecule of a specialized chlorophyll a which serves as an energy
trap. Photosystem II has a characteristic reactive centre namely a specialized chlorophyll-
protein complex. Photosystem I absorbs light at longer wavelength. Photosystem I1 is
activated by shorter wavelength, i.e. 670 nm and below and it is responsible for oxygen
evolution. Both the photosystems contain chlorophyll a and chlorophyll b. In
photosystem I, the ratio of chlorophyll a to chlorophyll b is higher than in photosystem
11'. These two photosystems must cooperate to yield maximum result in photosynthesis.

Chlorophyll a chlorophyll b
Vitamin B12
3. Enzymes
Enzymes are biological catalysts that govern, initiate and control biological reactivity
important for the life processes. They are produced by the living organism and are
usually present in only very small amounts in the various cells. All known enzymes are
proteins and some contain non-protein moieties termed prosthetic groups that are
essential for the manifestation of catalytic activities. In several natural enzymes,
metalloporphyrins constitute these prosthetic groups.

4. The Cytochromes
The cytochromes are electron transferring proteins, containing iron porphyrin, found in
aerobic cells. Some cytochromes found in endoplasmic reticulam, play a role in
specialized hydroxylation reactions. All cytochromes undergo reversible Fe (III)-Fe(II)
valency changes during their catalytic cycles. In almost all the cytochromes both the fifth
and sixth positions of the iron are occupied by the R groups of specific amino acid
residue of the proteins. Therefore, these cytochromes cannot bind with ligands like O 2,
CO or CN. An important exception is cytochrome oxidase that normally binds O2 in its
biological function.
The iron protoporphyrin group of cytochrome c is covalently linked to the protein by
thioether bridges between the porphyrin ring and two cysteine residue in the peptide
chain whereas in other cytochromes the porphyrin ring is non-covalently bound.
Cytochrome c is the only common heme moiety in which the heme is bound to the
protein by a covalent linkage.

Cytochromes
Applications of porphyrins and metalloporphyrins

The following are the applications of porphyrins and matalloporphyrins:

i. Porphyrins and related macrocycles provide an extremely versatile synthetic base for a
variety of material applications.
ii. porphyrins and metalloporphyrins have found applications as field-responsive materials,
Particularly for optoelectronic applications, including mesomorphic materials and
optical-limiting coatings.
iii. The porphyrin ligand serves as a platform on which one can erect desirable molecular and
materials properties.
iv. Porphyrins and metalloporphyrins can also be used as nonlinear optical materials.
v. They have desirable properties for use in optoelectronics.
vi. They have greater thermal stability and their extended π-conjugated macrocycle ring give
large nonlinear optical effects and subtle variation in their physical properties can be
made easily through chemical modification of their periphery.
vii. They also play key roles in adsorbing light energy over a wide spectral range and
converting it into the highly directional transfer of electrons.
viii. Porphyrins and metalloporphyrins are excellent candidates for a variety of sensing-
materials applications.
ix. They are also used in the detection of organic vapours and ionic species in solution.
x. Also metalloporphyrins exhibit very high photochemical stability.
LECTURE NOTE 3.

BIO-INORGANIC CHEMISTRY (CHEM. 411) (3 Credit Hours)

HEAVY METALS IN BIOLOGICAL SYSTEM AND ITS HEALTH EFFECTS

Metallic elements are intrinsic components of the environment. Their presence is considered
unique in the sense that it is difficult to remove them completely from the environment once they
enter in it. Metal constitute an important class of toxic substance which are encountered in
numerous occupational and environmental circumstances. The impact of these toxic agents on
human health is currently an area of intense interest due to the ubiquity of exposure. With the
increasing use of a wide verity of metals in industry and in our daily life, problems arising from
toxic metal pollution of the environment have assumed serious dimensions.

Sources and Emissions

Toxic metals, to a large extent, are dispersed in the environment through industrial effluents,
organic wastes, refuse burning, and transport and power generation. They can be carried to
places many miles away from the sources by wind, depending upon whether they are in gaseous
form or as particulates. Metallic pollutants are ultimately washed out of the air into land or the
surface of water ways. Thus air is also a route for the pollution of environment. Metal containing
industrial effluents constitute a major source of metallic pollution of hydrosphere. Another
means of dispersal is the movement of drainage water from catchment areas which have been
contaminated by waste from mining and smelting units. The chief toxic metals in industrial
effluents are shown in table:

METALS MANUFACTURING INDUSTRIES

Arsenic Phosphate and Fertilizer, Metal Hardening , Paints And Textile

cadmium Phosphate Fertilizer, Electronics, Pigments And Paints
chromium Metal Plating , Tanning, Rubber And Photography
copper Plating, Rayon And Electrical
Lead Paints, Battery
Nickel Electroplating , Iron Steel
Zinc Galvanizing, Plating Iron And Steel
Mercury Chlor-Alkali, Scientific Instruments , Chemicals
Toxic Effects

In general the toxicity of metal ions to mammalians systems is due to chemical reactivity of the
ions with cellular structural proteins, enzymes and membrane system. The target organs of
specific metal toxicities are usually those organs that accumulate the highest concentrations of
the metal in vivo. This is often dependent on the route of exposure and the chemical compound
of the metal i.e. its valiancy state, volatility, lipid solubility etc.

Besides the general toxicities of metals, we are today also concerned with the potential
carcinogenicity of metal compounds. Certain metals such as chromium and nickel have been
linked with cancers in exposed human populations. Metals have been shown to causes acute as
well as chronic poisoning in man and other experimental animals. Harmful effects of individual
metals are presented briefly below.

METAL TARGET ORGANS PRIMARY SOURCES CLINICAL

EFFECTS
Arsenic Pulmonary Nervous System, Industrial Dusts, Medicinal Perforation of nasal
Skin Uses Of Polluted Water septum Respiratory
Cancer, Peripheral
Neuropathy:
Dermatomes, Skin,
Cancer
Cadmium Renal, Skeletal Pulmonary Industrial Dust And Fumes Proteinuria,
And Polluted Water And Glucosuria,
Food Osteomalacia,
Aminoaciduria,
Emphysemi
Chromium Pulmonary Industrial Dust And Fumes Ulcer, Perforation of
And Polluted Food Nasal Septum,
Respiratory Cancer
Manganese Nervous System Industrial Dust And Fumes Central And
Peripheral
Neuropathies
Lead Nervous System, Industrial Dust And Fumes Encephalopathy,
Hematopoietic System, And Polluted Food Peripheral
Renal Neuropathy, Central
Nervous Disorders,
Anemia
Nickel Pulmonary, Skin Industrial Dust, Aerosols Cancer, Dramatis
Tin Nervous , Pulmonary Medicinal Uses, Industrial Central Nervous
System Dusts System Disorders,
Visual Defects And
EEG Changes,
 Pneumoconiosis.
Mercury Nervous System, Renal Industrial Dust And Fumes Proteinuria
And Polluted Water And
Food

Arsenic

Soluble inorganic arsenic can have immediate toxic effects. Ingestion of large amounts can lead
to gastrointestinal symptoms such as severe vomiting, disturbances of the blood and circulation,
damage to the nervous system, and eventually death. When not deadly, such large doses may
reduce blood cell production, break up red blood cells in the circulation, enlarge the liver, color
the skin, produce tingling and loss of sensation in the limbs, and cause brain damage

Long-term exposure to inorganic arsenic in drinking water in Taiwan has caused black foot
disease, in which the blood vessels in the lower limbs are severely damaged, resulting eventually
in progressive gangrene. The relationship between arsenic exposure and other health effects is
less clear. The evidence is strongest for high blood pressure, heart attacks and other circulatory
disease. The evidence is weaker for diabetes and reproductive effects; it is weakest for strokes,
longterm neurological effects, and cancer at sites other than lung, bladder, kidney and skin. as
well as other skin changes such as hyperkeratosis and pigmentation changes. These effects have
been demonstrated in many studies using different study designs. Exposure–response
relationships and high risks have been observed for each of these end-points. The effects have
been most thoroughly studied in Taiwan but there is considerable evidence from studies on
populations in other countries as well. Increased risks of lung and bladder cancer and of arsenic-
associated skin lesions have been reported to be associated with ingestion of drinking-water at
concentrations £ 50µg arsenic/litre.

Effects on human health Lead is a toxic heavy metal even at very low levels of exposure in
humans. Its effect on the human body can be both acute and chronic depending on dose and
exposure scenarios. Lead targets multiple organs in the body due to its systemic toxicity which
can cause neurological, cardiovascular, renal, gastro-intestinal, haematological and reproductive
effects. Human exposure to lead is usually tested through blood sampling. The lead stored in the
bones can emerge as a remobilised form of lead exposure late in the life of the individual.
Exposure occurs primarily through inhalation of dust particles, air contaminated with lead and
ingestion of foodstuffs, water and dust. Inhalation is an important exposure pathway for people
in the vicinity of point sources such as lead contaminated sites, countries where leaded fuel is
still used and areas where waste from products containing lead is burnt as well as secondary lead
recovery operations. Apart from ingestion of lead on foodstuffs and in drinking water a major
exposure source is leaded paint. Dust in homes containing leaded paint can be inhaled by adults
and children and ingested by children through pica behaviour.
Cadmium

Effects on human health Humans are exposed to cadmium by inhalation and ingestion although
the main health impacts recorded in the literature are through dietary exposure (kidney and bone
damage) and inhalation from smoking tobacco and occupational exposure (lung damage).
Dietary intake accounts for 90% of all exposure in non-smokers. Cadmium in the environment is
toxic to plants and animals and many micro-organisms. Cadmium does not degrade in the
environment to less toxic products which contributes to its bioaccumulation in the kidneys and
liver of vertebrates and invertebrates. Cadmium enters the environment from a variety of
anthropogenic sources. Wastewater is key source of environmental cadmium contamination and
diffuse pollution occurs through industrial air emissions and widespread use of fertilizers on
agricultural soils. Plants (including rice and tobacco) that are grown in contaminated soils take
up cadmium and lead to human dietary (and inhalation) exposures. However, human exposure
also occurs when cadmium contaminated soils are disturbed and the dust is inhaled. Diets high in
meat (especially liver and kidneys) or products from marine mammals may result in a
particularly high intake of cadmium.72 Cadmium is not considered essential for biological
function in humans. The main human organ impacted by cadmium exposure is the kidney in both
the general population and the occupationally exposed. Tobacco smokers are considered to be at
particular risk as are people with low iron levels. A secondary critical effect is skeletal damage
as a secondary response to kidney damage or direct action on the bone cells by the cadmium

Mercury

Main effects of Mercury on Human Health and the environment effects on human health
Toxicity of mercury is dependent on whether it takes the form of elemental mercury, inorganic
mercury or organic mercury compounds (particularly alkylmercury compounds such as
methylmercury and ethylmethyl salts and dimethylmercury). Accordingly, the exposure scenario
varies considerably for these different forms of mercury and complicates toxicity assessment. In
terms of methylmercury, dietary ingestion is the major source of human exposure, especially for
seafood and fish. Around 80% of inhaled elementary mercury vapour is retained in the tissue of
the lungs where it goes on to penetrate the blood-brain barrier where neurological effects take
place. Ingestion of elementary mercury does not always lead to high levels of absorption but
deaths have been reported. Inhalation of elementary mercury vapour has been observed to lead to
symptoms including tremors, emotional lability, insomnia, memory loss, neuromuscular changes,
and headaches as well effects on the kidney and thyroid. High exposures have led to death but
the critical effects are neurotoxic and renal.94 The main route of exposure to inorganic mercury
for humans is dietary although for some sub-sections of the population products such as skin
lightening creams, soaps and the use in traditional medicine and/ritualistic practices can result in
significant exposures to both inorganic an elemental mercury. Methylmercury is a well-known
potent neurotoxin which causes adverse impacts on the developing human brain. It passes readily
through the placental barrier and the blood-brain barrier making any exposure during pregnancy
of great concern. Methylmercury is considered possibly carcinogenic by the International
Agency for Research on Cancer.

Copper

Copper is a naturally-occurring metallic element that occurs in soil at an average concentration

of about 50 parts per million (ppm). It is present in all animals and plants and is an essential
nutrient for humans and animals in small amounts. The major sources of environmental copper
releases include the mining, smelting and refining of copper, industries producing products from
copper such as wire, pipes and sheet metal, and fossil fuel combustion. Water pipes are often
made of copper and bath fixtures may be made from brass and bronze alloys that contain copper.
The principal source of copper in drinking water results from the leaching of copper from pipes
and bath fixtures due to acidic water. Blue-green stains left in bath fixtures are a sign of the
presence of copper in water. Other releases of copper to the environment include agricultural use
against plant diseases and treatments applied to water bodies to eliminate algae. Health Effects
Absorption/Metabolism Studies investigating oral absorption of copper have found the
percentage absorbed ranging from 24-60 percent. Factors affecting the amount absorbed include
the amount of copper in the diet and competition with other metals found in food such as iron
and zinc.

Food sources rich in copper include shellfish, organ meats, nuts, beans and cocoa. Effects of
copper deficiency can include anemia, low numbers of white blood cells, osteoporosis in infants
and children, and defects in connective tissue leading to skeletal problems. Short-Term (Acute)
Effects Acute poisoning from ingestion of excessive copper can cause temporary gastrointestinal
distress with symptoms such as nausea, vomiting, and abdominal pain. Liver toxicity was seen in
doses high enough that resulted in death. High levels of exposure to copper can cause destruction
of red blood cells, possibly resulting in anemia. Long Term (Chronic) Effects Mammals have
efficient mechanisms to regulate copper stores in the body such that they are generally protected
from excess dietary copper levels. However, at high enough levels, chronic overexposure to
copper can damage the liver and kidneys. Wilson's disease is an inherited (genetic) disorder in
which copper builds up in the liver. Symptoms of liver toxicity (jaundice, swelling, pain) usually
do not appear until adolescence. Carcinogenicity (ability to cause cancer) although some studies
of workers exposed to copper have shown increased cancer risks, they were also exposed in the
workplace to other chemicals with carcinogenic potential. Increased cancer risk has not been
found in animal studies

Nickel

Nickel is a compound that occurs in the environment only at very low levels. Humans use nickel
for many different applications. The most common application of nickel is the use as an
ingredient of steal and other metal products. It can be found in common metal products such as
jewelry. Foodstuffs naturally contain small amounts of nickel. Chocolate and fats are known to
contain severely high quantities. Nickel uptake will boost when people eat large quantities of
vegetables from polluted soils. Plants are known to accumulate nickel and as a result the nickel
uptake from vegetables will be eminent. Smokers have a higher nickel uptake through their
lungs. Finally, nickel can be found in detergents. Humans may be exposed to nickel by breathing
air, drinking water, eating food or smoking cigarettes. Skin contact with nickel-contaminated soil
or water may also result in nickel exposure. In small quantities nickel is essential, but when the
uptake is too high it can be a danger to human health. An uptake of too large quantities of nickel
has the following consequences: Higher chances of development of lung cancer, nose cancer,
larynx cancer and prostate cancer Sickness and dizziness after exposure to nickel gas Respiratory
failure, Lung embolism, Birth defects, Asthma and chronic bronchitis, Allergic reactions such as
skin rashes, mainly from jewelry, Heart disorders. Nickel fumes are respiratory irritants and may
cause pneumonitis. Exposure to nickel and its compounds may result in the development of a
dermatitis known as “nickel itch” in sensitized individuals. The first symptom is usually itching,
which occurs up to 7 days before skin eruption occurs. The primary skin eruption is
erythematous, or follicular, which may be followed by skin ulceration. Nickel sensitivity, once
acquired, appears to persist indefinitely.

Tin

Scientists use many tests to protect the public from harmful effects of toxic chemicals and to find
ways for treating persons who have been harmed. One way to learn whether a chemical will
harm people is to determine how the body absorbs, uses, and releases the chemical. For some
chemicals, animal testing may be necessary. Animal testing may also help identify health effects
such as cancer or birth defects. Without laboratory animals, scientists would lose a basic method
for getting information needed to make wise decisions that protect public health. Scientists have
the responsibility to treat research animals with care and compassion. Scientists must comply
with strict animal care guidelines because laws today protect the welfare of research animals.
Because inorganic tin compounds usually enter and leave your body rapidly after you breathe or
eat them, they do not usually cause harmful effects. However, humans who swallowed large
amounts of inorganic tin in research studies suffered stomachaches, anemia, and liver and kidney
problems. Studies with inorganic tin in animals have shown similar effects to those observed in
humans. There is no evidence that inorganic tin compounds affect reproductive functions,
produce birth defects, or cause genetic changes. Inorganic tin compounds are not known to cause
cancer. Inhalation (breathing in), oral (eating or drinking), or dermal exposure (skin contact) to
some organotin compounds has been shown to cause harmful effects in humans, but the main
effect will depend on the particular organotin compound. There have been reports of skin and
eye irritation, respiratory irritation, gastrointestinal effects, and neurological problems in humans
exposed for a short period of time to high amounts of certain organotin compounds. Some
neurological problems have persisted for years after the poisoning occurred. Lethal cases have
been reported following ingestion of very high amounts.

Lead

Most of the studies looking for a possible link between lead exposure and cancer have focused
on workers with high levels of occupational (work-related) exposure to inorganic lead. People
with heavy workplace exposures to lead have been found to have blood lead concentrations
many times higher than the average blood lead concentration in the general population. Several
studies have looked for a link between exposure to lead in the workplace (mainly among battery
workers and smelter workers) and lung cancer. Some of these studies have found a small
increase in lung cancer risk. However, most of these studies were limited in that they didn't take
into account other factors that might affect lung cancer risk, such as smoking or exposures to
arsenic or other heavy metals that typically also occur along with lead exposures in industrial
settings. Some studies looking at blood lead levels in the general population have also found a
small increased risk of lung cancer in people with higher lead levels. Several of these same
workplace studies also looked at stomach cancer risk. Most of the studies found an increased risk
of stomach cancer with higher lead exposure. Although it is unlikely these results would be
affected by smoking or arsenic exposure, the studies didn’t take into account other factors that
could also have affected stomach cancer risk. Studies have also looked at possible links between
workplace exposures to lead and other cancers, including cancers of the brain, kidney, bladder,
colon, and rectum. The results of these studies have been mixed. Some studies have found links,
while others have not. The link between lead exposure and cancer is clearly a concern, and more
research is needed to better define the possible link between lead exposure and a number of
cancers

LECTURE NOTE 4.

BIO-INORGANIC CHEMISTRY (CHEM. 411) (3 Credit Hours)

CHEMICAL TOXICOLOGY

Definition of toxicology: This is study of the adverse effects of chemicals or physical agents on
living organisms.

The traditional definition of toxicology is "the science of poisons." As our understanding of how
various agents can cause harm to humans and other organisms, a more descriptive definition of
toxicology is "the study of the adverse effects of chemicals or physical agents on living
organisms”.

Adverse effects may occur in many forms, ranging from immediate death to subtle changes not
realized until months or years later. They may occur at various levels within the body, such as an
organ, a type of cell, or a specific biochemical. Knowledge of how toxic agents damage the body
has progressed along with medical knowledge. It is now known that various observable changes
in anatomy or body functions actually result from previously unrecognized changes in specific
biochemical in the body.

Toxicology Terminology

1. A toxicologist: This is a scientist that determines the harmful effects of agents and the cellular,
biochemical, and molecular mechanisms responsible for the effects.

2. Toxicants: This is substance that produce adverse biological effect of any nature.

3. Toxin: specific protein produce by living organisms, most exhibit immediate effects.

4. Poisons: toxicants that cause immediate death or illness when experienced in a very small
amount.

Toxicant, toxin, and poison are often used interchangeably in the literature; however, there are
subtle differences as indicated in the above. Toxic substances may be systemic toxins or organ
toxins. A systemic toxin is one that affects the entire body or many organs rather than a specific
site. For example, potassium cyanide is a systemic toxicant in that it affects virtually every cell
and organ in the body by interfering with the cell's ability to utilize oxygen. Toxicants may also
affect only specific tissues or organs while not producing damage to the body as a whole. These
specific sites are known as the target organs or target tissues. Some examples: Benzene is a
specific organ toxin in that it is primarily toxic to the blood-forming tissues. Lead is also a
specific organ toxin; however, it has three target organs (central nervous system, kidney, and
hematopoietic system).

A toxic agent
This is anything that can produce an adverse biological effect. It may be chemical, physical, or
biological in form. For example, toxic agents may be chemical (such as cyanide), physical (such
as radiation) and biological (such as snake venom).

A distinction is made for diseases due to biological organisms. Those organisms that invade and
multiply within the organism and produce their effects by biological activity are not classified as
toxic agents. An example of this is a virus that damages cell membranes resulting in cell death.

If the invading organisms excrete chemicals which is the basis for toxicity, the excreted
substances are known as biological toxins. The organisms in this case are referred to as toxic
organisms. An example is tetanus. Tetanus is caused by a bacterium, Clostridium tetani. The
bacteria C. tetani itself does not cause disease by invading and destroying cells. Rather, it is a
toxin that is excreted by the bacteria that travels to the nervous system (a neurotoxin) that
produces the disease.

A toxic substance

This is simply a material which has toxic properties. It may be a discrete toxic chemical or a
mixture of toxic chemicals. For example, lead chromate, asbestos, and gasoline are all toxic
substances. Lead chromate is a discrete toxic chemical. Asbestos is a toxic material that does not
consist of an exact chemical composition but a variety of fibers and minerals. Gasoline is also a
toxic substance rather than a toxic chemical in that it contains a mixture of many chemicals.
Toxic substances may not always have a constant composition. For example, the composition of
gasoline varies with octane level, manufacturer, time of season, etc. oxic substances may be
organic or inorganic in composition.

Organic Toxins: Substance that were originally derived from

living organisms (thus named organic).

Inorganic Toxins: specific chemicals that were not derived from

living organisms (chemical), generally few molecules consisting of
only a small atoms such as nitrogen.

Toxic substances may be systemic toxins or organ toxins. A systemic toxin is one that affects the
entire body or many organs rather than a specific site. For example, potassium cyanide is a
systemic toxicant in that it affects virtually every cell and organ in the body by interfering with
the cell's ability to utilize oxygen.

Toxicants may also affect only specific tissues or organs while not producing damage to the
body as a whole. These specific sites are known as the target organs or target tissues.
Examples: Benzene is a specific organ toxin in that it is primarily toxic to the blood-forming
tissues. Lead is also a specific organ toxin; however, it has three target organs (central nervous
system, kidney, and hematopoietic system).

A toxicant may affect a specific type of tissue (such as connective tissue) that is present in
several organs. The toxic site is then referred to as the target tissue. There are many types of cells
in the body and they can be classified in several ways. basic structure (e.g., cuboidal cells) tissue
type (e.g., hepatocytes of the liver) germinal cells (e.g., ova and sperm) somatic cells (e.g., non-
reproductive cells of the body)
Factors determining adverse effects
Germ cells are those cells that are involved in the reproductive process and can give rise to a new
 intrinsic
organism. They toxicity
have only a single set of chromosomes peculiar to a specific sex. Male germ
 dose
cells give rise to sperm and female germ cells develop into ova. Toxicity to germ cells can cause
 developing
effects on the exposure conditions
fetus (such as birth defects, abortions). Somatic cells are all body cells
 response of
except the reproductive germ host
cells. They have two sets (or pairs) of chromosomes. Toxicity to
somatic cells causes a variety of toxic effects to the exposed individual (such as dermatitis,
death, and cancer).

Dose by definition is the amount of a substance administered at one time. However, other
parameters are needed to characterize the exposure to xenobiotics. The most important are the
number of doses, frequency, and total time period of the treatment.

Some examples: 500 mg Asperin as a single dose, 500 mg Penicillin every 8 hours for 10 days,
15 mg DDT per day for 60 days

Types of doses

1. Exposure dose: the amount of xenobiotic encountered in the environment.

2. Absorbed dose: the actual amount of the exposes dose that enters the body
3. Administered dose: the quantity administered usually orally or by injection

4. Total dose: the sum of all the individual doses.

Fractionating a total dose usually decreases the probability that the total dose will cause toxicity.
The reason for this is that the body often can repair the effect of each sub-toxic dose if sufficient
time passes before receiving the next dose. In such a case, the total dose, harmful if received all
at once, is non-toxic when administered over a period of time. For example, 30 mg of strychnine
swallowed at one time could be fatal to an adult whereas 3 mg of strychnine swallowed each day
for ten days would not be fatal.

DOSE-RESPONSE RELATIONSHIP

The dose-response relationship is a fundamental and essential concept in toxicology. It correlates

exposures and the spectrum of induced effects. Generally, the higher the dose, the more severe
the response. The dose-response relationship is based on observed data from experimental
animal, human clinical, or cell studies.

Dose

Knowledge of the dose-response relationship: - establishes causality that the chemical has in fact
induced the observed effects - establishes the lowest dose where an induced effect occurs - the
threshold effect - determines the rate at which injury builds up - the slope for the dose response.

Within a population, the majority of responses to a toxicant are similar; however, a wide
variance of responses may be encountered, some individuals are susceptible and others resistant.
As demonstrated above, a graph of the individual responses can be depicted as a bell shaped
standard distribution curve.

Dose responses are commonly presented as mean + 1 S.D. (standard deviation), which
incorporates 68% of the individuals. The variance may also be presented as two standard
deviations, which incorporates 95% of the responses. A large standard deviation indicates great
variability of response. For example, a response of 12+5 mg indicates considerably more
variability than 12+1 mg.

Dose

The dose-response curve normally takes the form of a sigmoid curve. It conforms to a smooth
curve as close as possible to the individual data points. For most effects, small doses are not
toxic. The point at which toxicity first appears is known as the threshold dose level. From that
point, the curve increases with higher dose levels. In the hypothetical curve above, no toxicity
occurs at 10 mg whereas at 35 mg 100% of the individuals experience toxic effects.

There is always a relation between dose and effect/response, but for some agents there is a
threshold below which no effect occurs. A threshold for toxic effects occurs at the point where
the body's ability to detoxify a xenobiotic or repair toxic injury has been exceeded. For most
organs there is a reserve capacity so that loss of some organ function does not cause decreased
performance. For example, the development of cirrhosis in the liver may not result in a clinical
effect until over 50% of the liver has been replaced by fibrous tissue.
 Dose

Knowledge of the shape and slope of the dose-response curve is extremely important in
predicting the toxicity of a substance at specific dose levels. Major differences among toxicants
may exist not only in the point at which the threshold is reached but also in the percent of
population responding per unit change in dose (i.e., the slope). As illustrated above, Toxicant A
has a higher threshold but a steeper slope than Toxicant B.

Knowledge of the dose-response relationship permits one to determine whether exposure has
caused an effect, threshold for the effect, and the rate of buildup of the effect with increasing
dose levels. Rate of buildup of toxic effects is known as the "slope" of the dose-response curve.
 Dose estimates

Dose Estimates of Toxic Effects

Dose-response curves are used to derive dose estimates of chemical substances. A common dose
estimate for acute toxicity is the LD50 (Lethal Dose 50%). This is a statistically derived dose at
which 50% of the individuals will be expected to die. The figure illustrates how an LD50 of 20
mg is derived. Other dose estimates also may be used. LD0 represents the dose at which no
individuals are expected to die. This is just below the threshold for lethality. LD10 refers to the
dose at which 10% of the individuals will die.

For inhalation toxicity, air concentrations are used for exposure values. Thus, the LC50 is
utilized which stands for Lethal Concentration 50%, the calculated concentration of a gas lethal
to 50% of a group. Occasionally LC0 and LC10 are also used.

Effective Doses (EDs) are used to indicate the effectiveness of a substance. Normally, effective
dose refers to a beneficial effect (relief of pain). It might also stand for a harmful effect. Thus the
specific endpoint must be indicated.

Toxic Doses (TDs) are utilized to indicate doses that cause adverse toxic effects. The usual dose
estimates are seen in the table. The knowledge of the effective and toxic dose levels aides the
toxicologist and clinician in determining the relative safety of toxic agents. Two dose-response
curves can be presented for the same drug, one for effectiveness and the other for toxicity. In this
case, a dose that is 50-75% effective does not cause toxicity whereas a 90% effective dose may
result in a small amount of toxicity.
The toxic dose is seen at an increasing dose compared with the effective dose. As can been seen
in the graph, it is ppossible to have an overlap of having an effective dose while this is also
starting to be toxic.

Question: The quantity of a substance administered to an individual over a period of time or in

several individual doses is known as the:

 Exposure Dose
 Absorbed Dose
 Total Dose

Answer: The total dose is the quantity of a substance administered to an individual over a period
of time or in several individual doses. It becomes particularly important when evaluating
cumulative poisons.
 NOAEL and LOAEL

Two terms often encountered are No Observed Adverse Effect Level (NOAEL) and Low
Observed Adverse Effect Level (LOAEL). They are the actual data points from human clinical
or experimental animal studies.

Sometimes the terms No Observed Effect Level (NOEL) and Lowest Observed Effect Level
(LOEL) may also be found in the literature. NOELs and LOELs do not necessarily imply toxic
or harmful effects and may be used to describe beneficial effects of chemicals as well.

The NOAEL, LOAEL, NOEL, and LOEL have great importance in the conduct of risk
assessments.

Toxic effects

A target organ is an organ that is damaged by the

xenobiotic or its metabolite. There may be more than
one target for toxicity for a particular substance.

For example, the targets for alcohol are the central

nervous system and the liver.

Toxicity is complex with many influencing factors; dosage is the most important. Xenobiotics
cause many types of toxicity by a variety of mechanisms. Some chemicals are themselves toxic.
Others must be metabolized (chemically changed within the body) before they cause toxicity.
Many xenobiotics distribute in the body and often affect only specific target organs. Others,
however, can damage any cell or tissue that they contact. The target organs that are affected may
vary depending on dosage and route of exposure. For example, the target for a chemical after
acute exposure may be the nervous system, but after chronic exposure the liver.

Toxicity can result from adverse cellular, biochemical, or macromolecular changes.

Examples are:

 cell replacement, such as fibrosis

 damage to an enzyme system
 disruption of protein synthesis
 production of reactive chemicals in cells
 DNA damage

Some xenobiotics may also act indirectly by:

 modification of an essential biochemical function

 interference with nutrition
 alteration of a physiological mechanism

SYSTEMIC TOXIC EFFECTS

Toxic effects are generally categorized according to the site of the toxic effect. Sometimes the
effect may occur at only one site. This site is referred to as the specific target organ. In other
cases, toxic effects may occur at multiple sites. This is referred as systemic toxicity. Following
are types of systemic toxicity:

i. Acute Toxicity
ii. Subchronic Toxicity
iii. Chronic Toxicity
iv. Carcinogenicity Developmental Toxicity
v. Genetic Toxicity (somatic cells)

Acute toxicity occurs almost immediately (hours/days) after an exposure. An acute exposure is
usually a single dose or a series of doses received within a 24 hour period. Death is a major
concern in cases of acute exposures. Examples, Many people die each year from inhaling carbon
monoxide from faulty heaters. Non-lethal acute effects may also occur, e.g., convulsions and
respiratory irritation.
Subchronic Toxicity: Subchronic toxicity results from repeated exposure for several weeks or
months. This is a common human exposure pattern for some pharmaceuticals and environmental
agents. Examples are: Ingestion of coumadin tablets (blood thinners) for several weeks as a
treatment for venous thrombosis can cause internal bleeding, Workplace exposure to lead over a
period of several weeks can result in anemia.

Chronic Toxicity: Chronic toxicity represents cumulative damage to specific organ systems and
takes many months or years to become a recognizable clinical disease. Damage due to
subclinical individual exposures may go unnoticed. With repeated exposures or long-term
continual exposure, the damage from these subclinical exposures slowly builds-up (cumulative
damage) until the damage exceeds the threshold for chronic toxicity. Ultimately, the damage
becomes so severe that the organ can no longer function normally and a variety of chronic toxic
effects may result. Examples of chronic toxic effects are:

 cirrhosis in alcoholics who have ingested ethanol for several years;

 chronic kidney disease in workmen with several years exposure to lead;
 chronic bronchitis in long-term cigarette smokers;
 Pulmonary fibrosis in coal miners (black lung disease).

Developmental Toxicity: Developmental Toxicity pertains to adverse toxic effects to the

developing embryo or fetus. This can result from toxicant exposure to either parent before
conception or to the mother and her developing embryo-fetus.

Chemicals cause developmental toxicity by two methods. They can act directly on cells of the
embryo causing cell death or cell damage, leading to abnormal organ development. A chemical
might also induce a mutation in a parent's germ cell which is transmitted to the fertilized ovum.
Some mutated fertilized ova develop into abnormal embryos.

Carcinogenicity is a complex multistage process of abnormal cell growth and differentiation

which can lead to cancer. At least two stages are recognized. They are initiation in which a
normal cell undergoes irreversible changes and promotion in which initiated cells are stimulated
to progress to cancer. Chemicals can act as initiators or promoters.

The initial neoplastic transformation results from the mutation of the cellular genes that control
normal cell functions. The mutation may lead to abnormal cell growth. It may involve loss of
suppresser genes that usually restrict abnormal cell growth. Many other factors are involved
(e.g., growth factors, immune suppression, and hormones).

A tumor (neoplasm) is simply an uncontrolled growth of cells. Benign tumors grow at the site of
origin; do not invade adjacent tissues or metastasize; and generally are treatable. Malignant
tumors (cancer) invade adjacent tissues or migrate to distant sites (metastasis). They are more
difficult to treat and often cause death.

Genetic Toxicity: Genetic Toxicity results from damage to DNA and altered genetic expression.
This process is known as mutagenesis. The genetic change is referred to as a mutation and the
agent causing the change as a mutagen. There are three types of genetic change

If the mutation occurs in a germ cell the effect is heritable. There is no effect on the exposed
person; rather the effect is passed on to future generations. If the mutation occurs in a somatic
cell, it can cause altered cell growth (e.g. cancer) or cell death (e.g. teratogenesis) in the exposed
person.

NOTE.

ASSIGNMENT

Make a comprehensive note on Chemical carcinogens and harmful drugs.

To be submitted one week before the first semester examination